Primary open angle glaucoma (POAG), the most common form of glaucoma, is usually accompanied by elevated intraocular pressure (IOP) due to failure of the trabecular meshwork (TM) to maintain normal levels of aqueous humor outflow. We recently generated a novel transgenic murine model (Tg-MYOCY437H) that expresses mutant myocilin, a known leading genetic cause of POAG in humans and replicates human glaucoma phenotypes. Importantly, we have associated endoplasmic reticulum (ER) stress to the pathogenesis of glaucoma in Tg-MYOCY437H mice. Misfolded myocilin accumulates in the ER, induces ER stress and activates a protective unfolded protein response (UPR). Along with UPR activation, mutant myocilin also induces autophagy, a process of lysosomal degradation known to degrade protein aggregates. However, failure to eliminate myocilin aggregates possibly due to insufficient UPR and impaired autophagy, TM cells induce the ER stress-initiated apoptotic transcriptional factor, Chop, which may further worsen ER homeostasis and cause TM dysfunction/loss, elevating IOP and resulting in POAG. The current proposal will further investigate the role of chronic ER stress in the pathogenesis of myocilin as well as non-myocilin associated POAG. During the mentored phase, the proposed studies will determine whether failure to activate the protective UPR (Atf-6-/-) exacerbates glaucoma phenotypes, whereas interference with ER stress-induced apoptosis (Chop-/-) prevents glaucoma in Tg-MYOCY437H mice. In addition, we will investigate whether ER stress is activated in port-mortem TM tissues from POAG donors. During the independent phase, we will examine whether induction of ER stress in the TM is associated with elevation of IOP in a mouse model of dexamethasone-induced ocular hypertension. In addition, we will examine the role of autophagy in degradation of myocilin aggregates and will determine whether inducing autophagy by rapamycin will rescue the glaucoma of Tg-MYOCY437H mice. Specifically, the candidate will learn and generate new models of ER stress using Atf6 and Chop knockout mice, generate induced pluripotent stem cells (iPSCs)-derived trabecular meshwork- like cells from POAG patients, and characterize a dexamethasone-induced ocular hypertension mouse model in the laboratory of Dr. Val Sheffield. Additionally, during the mentored phase, the candidate will continue his professional and scientific career development through continual guidance from the advisory committee. He will attend scientific conferences, and collaborate with ER stress expert, Dr. Thomas Rutkowski and glaucoma clinical expert, Dr. Lee Alward. He will also acquire teaching experience. This project will facilitate continued technical, intellectul, and professional training of the candidate, and assist the candidate in the establishment of an independent research laboratory at an academic research institute. PUBLIC HEALTH RELEVANCE: The successful completion of this project will lead to insight into elevated intraocular pressure, a major risk factor of the human blinding disorder, primary open angle glaucoma. The results will potentially improve diagnosis, genetic risk assessment and treatment of this common disorder.